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Cell Biology JoVE (Open Access) Citable · DOI

Metabolic Profile Analysis of Zebrafish Embryos

DOI: 10.3791/4300-v
What you'll learn
  • ✓ Measure in vivo metabolic profiles of zebrafish embryos using Seahorse XF analyzer
  • ✓ Compare mitochondrial function parameters between genetically or pharmacologically manipulated samples
  • ✓ Perform chemical treatment and live metabolic analysis on developing zebrafish
  • ✓ Analyze lipid accumulation via Oil Red O staining and correlate with metabolic data
Protocol

Biopharma Insights Zebrafish represent a powerful vertebrate model that has been under-utilised for metabolic studies. Here we describe a rapid way to measure the in vivo metabolic profile of developing zebrafish that allows the comparison of different mitochondrial function parameters between genetically or pharmacologically manipulated embryos, thereby increasing the applicability of this organism.

Difficulty
intermediate
Total time
~4–6 hours per experiment (embryo treatment, metabolic measurement, and staining)
Model organism
Zebrafish (Danio rerio) embryos
Biosafety
BSL-1

Steps

1
Chemically treat zebrafish embryos and prepare for analysis

Apply chemical treatments or genetic manipulations to developing zebrafish embryos. Prepare treated embryos for live metabolic profiling on the Seahorse XF analyzer platform.

▶ 01:23
2
Measure live metabolic profile using Seahorse XF analyzer

Load treated embryos into Seahorse XF analyzer cartridges and perform real-time measurement of oxygen consumption rate and extracellular acidification rate to assess mitochondrial function parameters.

▶ 02:38
3
Analyze metabolic data and perform Oil Red O staining

Process and analyze Seahorse XF output data to compare mitochondrial parameters across samples. Perform Oil Red O staining to visualize and quantify lipid accumulation in embryos.

▶ 04:40
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